【摘要】 目的:建立大鼠胚胎视网膜祖细胞体外培养方法并观察其分化特性。方法:取孕18d大鼠胚胎,用无血清培养技术扩增视网膜祖细胞;用免疫细胞化学、扫描电镜与透射电镜技术对祖细胞的特性及诱导分化的细胞类型进行鉴定。结果:从胚胎大鼠视网膜神经部分离的细胞具有增殖能力,可进行传代培养,获得的细胞球能表达Nestin并具有掺入BrdU的能力。诱导分化后的细胞表达,其中,Thy1.1+细胞最多,Opsin+细胞次之,GS+细胞较少,PKC+细胞少,Syntaxin+及Peripherin+细胞极少,不表达CalbindinD28K。结论:从胚胎大鼠分离的细胞具有自我增殖能力和多向分化潜能,属于视网膜祖细胞;诱导分化产生5种视网膜细胞,以节细胞最多,未见水平细胞。更多还原

【Abstract】 Objective: To isolate and incubate neural progenitor cells from rat embryonic retina and to explore their characteristics of differentiation.Methods: The neural progenitor cells were isolated from rat embryonic retina by using serum-free culture, and the cells were cultured and passaged in vitro. Immunocytochemistry, transmission and scanning electron microscopy were used to identify retinal progenitor cells and their potentiality of differentiation. Results: The cells isolated from rat embryonic retina possessed the ability of proliferation and had been passaged in vitro. They formed into spheres, expressed nestin and was BrdU-positive after incubation in the medium with BrdU. After induced-differentiation, they expressed many retinal cell-typical markers but Calbindin D-28K. Differentiated cells were antigen-positive with most being Thy1.1 +cells, then Opsin + cells, few GS + cells, very few PKC +, Syntaxin + and Peripherin + cells.Conclusion: The cells harvested from retina in vitro have the ability of self-proliferation and multipotential differentiation. These cells can generate 5 types of retinal cells most of which are ganglion cells.更多还原